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Separating and purifying method for preparing white muscardine silkworm anticancer-activity polysaccharide BBPW-1

A technology of BBPW-1 and anti-cancer activity, which is applied in the field of separation, purification and preparation of BBPW-1 anti-cancer active polysaccharide of Bombyx botany problems, to achieve the effect of improving medicinal value, positive social and economic benefits, and uniform purity

Inactive Publication Date: 2013-08-28
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the medicinal mechanism of silkworm, especially the anti-cancer mechanism, is still unclear, which hinders the improvement of the medicinal level and the expansion of the medicinal scope of silkworm

Method used

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  • Separating and purifying method for preparing white muscardine silkworm anticancer-activity polysaccharide BBPW-1
  • Separating and purifying method for preparing white muscardine silkworm anticancer-activity polysaccharide BBPW-1

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Take the silkworm powder passed through an 80-mesh sieve, degrease, remove glycosides and alkaloids according to the above method, add distilled water at a ratio of 1:5 to liquid, treat with ultrasonic waves for 40 min, extract at 100°C for 1 h, and extract Crude polysaccharides; crude polysaccharides were deproteinized by the Sevag method (volume ratio chloroform: n-butanol = 4:1), put on a DEAE-Sepharose Fast Flow ion-exchange chromatography column, eluted with distilled water at a flow rate of 3.0 mL / min, and collected automatically The neutral polysaccharide component BBPW was obtained; BBPW was placed on a Sephacryl S-300 chromatography column, eluted with distilled water at a flow rate of 1.0 mL / min, collected by an automatic collector, and the eluate of the first elution peak was collected, and reduced Concentrate under reduced pressure; put the concentrated solution on a Sephacryl S-500 chromatography column, elute with distilled water at a flow rate of 1.0 mL / mi...

Embodiment 2

[0022] Take the silkworm powder passed through a 100-mesh sieve, degrease, remove glycosides and alkaloids according to the above method, add distilled water at a ratio of 1:5 to liquid, treat with ultrasonic waves for 50 min, extract at 100°C for 1.5 h, and extract Crude polysaccharides; crude polysaccharides were deproteinized by the Sevag method (volume ratio chloroform: n-butanol = 4:1), put on a DEAE-Sepharose Fast Flow ion-exchange chromatography column, eluted with distilled water at a flow rate of 3.5 mL / min, and collected automatically The neutral polysaccharide component BBPW was obtained; the Sephacryl S-300 chromatographic column on BBPW was eluted with distilled water at a flow rate of 1.2 mL / min, collected by an automatic collector, and the eluate of the first elution peak was collected, and reduced concentrated under reduced pressure; the concentrated solution was put on a Sephacryl S-500 chromatography column, eluted with distilled water at a flow rate of 1.2 mL...

Embodiment 3

[0024] Take the silkworm powder passed through a 90-mesh sieve, degrease, remove glycosides and alkaloids according to the above method, add distilled water at a ratio of 1:5 to liquid, treat with ultrasonic waves for 40 min, extract at 100°C for 1.2 h, and extract Crude polysaccharides; crude polysaccharides were deproteinized by the Sevag method (volume ratio chloroform: n-butanol = 4:1), put on a DEAE-Sepharose Fast Flow ion-exchange chromatography column, eluted with distilled water at a flow rate of 3.2 mL / min, and collected automatically The neutral polysaccharide component BBPW was obtained; the Sephacryl S-300 chromatographic column on BBPW was eluted with distilled water at a flow rate of 1.1 mL / min, collected by an automatic collector, and the eluate of the first elution peak was collected, and reduced Concentrate under reduced pressure; put the concentrated solution on a Sephacryl S-500 chromatography column, elute with distilled water at a flow rate of 1.1 mL / mi...

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Abstract

The invention relates to a separating and purifying method for preparing white muscardine silkworm anticancer-activity polysaccharide BBPW-1, which comprises the following steps: degreasing white muscardine silkworm powder under reflux of acetone-petroleum ether and 80% ethanol to remove glycosides and alkaloids, and extracting with distilled water at 80-100 DEG C to obtain crude polysaccharide; and after removing proteins from the crude polysaccharide by a Sevag process, separating out the neutral polysaccharide component by DEAE (diethylaminoethanol) sepharose ion-exchange chromatography, carrying out propylene dextrangel S-300 chromatography, taking the first eluting peak, concentrating under reduced pressure, carrying out propylene dextrangel S-500 chromatography for further purification, and carrying out freeze-drying to obtain the white muscardine silkworm anticancer-activity polysaccharide BBPW-1. The product prepared by the method provided by the invention has uniform purity, and the molecular weight is 3.67*10<6>Da; and the product has an inhibiting action on growth of human cervical cancer cells Hela and human liver cancer cells HepG2, does not have any adverse effect on growth of normal human embryo kidney cells HEK293 and mouse macrophages RAW264.7, and can be used for developing anticancer products.

Description

technical field [0001] The invention relates to a method for separating, purifying and preparing anticancer active polysaccharide BBPW-1 from Bombyx mori. Background technique [0002] Natural polymer polysaccharides, mainly from higher animal and plant cell membranes and microbial cell walls, are one of the four basic substances that constitute life activities. Polysaccharides have a variety of biological activities. Glycoproteins formed with proteins and lipopolysaccharides formed with lipids play an important role in cell recognition, secretion, and protein processing and transport. Since mycopolysaccharides were found to have anti-tumor activity in the late 1950s, a variety of anti-cancer polysaccharides have been extracted from bacteria, fungi, algae, plants and animals and used clinically. Due to the effective anti-cancer effect of natural polysaccharides and low Toxicity has been paid more and more attention by people. [0003] Bombyx mori is a silkworm ( Bombyx mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/00A61P35/00
Inventor 时连根蒋学
Owner ZHEJIANG UNIV
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